The present invention is related to devices and methods of using the same, to prepare pharmaceutical compositions comprising a complex of an antigen component and the corresponding antibody or inhibitor component in a pharmacologically acceptable carrier. Specifically, the invention concerns a device and method of using the same, of preparing a pharmaceutical composition by selectively isolating an antibody type component from a physiological fluid, such as blood, plasma or other bodily fluid in a pharmacologically acceptable carrier to which the antigen type compound may be added.
Pharmaceutical compositions containing antibody/antigen complexes have been suggested for numerous medical applications. For example, U.S. Pat. No. 4,740,371, issued to St. Remy et al on Apr. 26, 1988, disclosed a complex useful for treating allergies. This complex includes the specific allergen that causes the allergic reaction, and the corresponding antibody for that allergen. While the antibodies may be derived from either the patient's or a donor's physiological fluids, the preferred antibody is usually derived from the patient's own physiological fluids. The injection of this complex reduces, and even eliminates, a patient's allergic reaction to the specific allergen, while not possessing the side effects accompanying conventional allergy treatments.
Another antibody/antigen complex is taught in EP 426913, published May 15, 1991. This application discloses a complex composed of factor VIII and an antibody termed factor VIII inhibitor. This complex is used to treat hemophiliacs who have become refractory to the injection of factor VIII. These individuals produce a factor VIII inhibitor antibody which binds with, and renders factor VIII inactive. Thus factor VIII functions as the antigen, while the anti-factor VIII functions as the antibody. It was demonstrated that the injection of the factor VIII antigen/inhibitor complex reduces some patient's refractortness to factor VIII injections.
The antigen/antibody complexes taught in St. Remy et al and the pending application preferably use the antibody from the patient's own physiological fluids. This is significant not only because it eliminates any potential reaction to an antibody prepared from a non-identical donor, but more importantly, autologous antibodies are most effective for induction of the therapy itself. The presently available methods, as taught by St. Remy et al and the referenced application, of extracting the antibodies requires painstaking chemical isolation of the antibody from the patient's bodily fluid, such as blood. For example, St. Remy et al discloses isolating the desired antibodies IgG, IgM, IgA, IgE, and IgD by a complex series of steps involving precipitation, dialysis, concentration, chromatography and immunoadsorption.
Thus while pharmaceutical compositions containing antigen/antibody complexes may be beneficial in the treatment of certain diseases and conditions, the methods of isolating the desired antibody are complex and time consuming. It is thus desirable to provide a mechanism to readily prepare pharmaceutical antibody/antigen complexes.
Recent advances in separation technology have presented the opportunity of isolating a specific target population, e.g. cells, proteins, or antibodies without the need of tedious and extensive chemical separation techniques. For example, various workers have suggested capturing specific target populations by using filters bearing immunoreactive groups. Immunoreactive groups are those to which the desired antibody will selectively bind, typically the antigen. These techniques include filters prepared from fibers bearing immunoreactive groups, U.S. Pat. No. 3,843,324, issued on Oct. 22, 1974; columns bearing immunoreactive group, U.S. Pat. No. 4,252,653, issued on Feb. 24, 1981; and filter candles bearing immunoreactive groups, U.S. Pat. No. 4,648,974, issued Mar. 10, 1987.
Another recently developed technique for isolating target populations, i.e. antibodies, selected proteins and cells, from a physiological fluid utilizes paramagnetic beads or particles coated with an immunoreactive compound or agent selective for the desired target population. Examples of such particles or beads are disclosed in U.S. Pat. Nos. 4,230,685, issued Oct. 28, 1980; 4,554,088, issued Nov. 19, 1985; and 4,628,037, issued Dec. 9, 1986. The use of such particles in the separation of taught in publications, "Removal of Neuroblastoma Cells From Bone Marrow with Monoclonal Antibodies Conjugated to Magnetic Microspheres", by J. G. Treleaven, J. Ugelstad, T. Philips, F. M. Gibson, A. Rembaum, G. D. Caines and J. T. Kemshead, The Lancet, Jan. 14, 1984, pages 70-73, and "Immunomagnetic removal of B-lymphoma cells from human bone marrow: a procedure for clinical use", by G. Kvalheim, O. Sorensen, O. Fodstad, S. Funderud, S. Kiesel, B. Dorken, K. Nustad, E. Jakobsen, O. Ugelstad and A, Pihl, Bone Marrow Transplantation, (1988), volume 3, pages 31-41.
Other references which disclose devices and methods of isolating specific target populations are U.S. Pat. Nos. 3,970,518, and 4,018,886 both issued to Giaever on Jul. 20, 1976 and Apr. 19, 1977, respectively, 4,219,411, issued to Yen et al on Aug. 26, 1980 and 4,710,472, issued to Saur et al on Dec. 1, 1987. Both Yen et al and Saur et al disclose intricate devices for mixing a physiological fluid with Immunoreactive paramagnetic particles. These immunoreactive paramagnetic particles are coated with an agent specific for the target population, that is, will selectively bind to the target population cellular members or chemical species. These paramagnetic particles are subsequently captured by a magnetic field while removing the fluid.
The Giaever patents disclose devices and methods of isolating the select population of cells from a physiological fluid using the immunoreactive paramagnetic particles. The immunoreactive paramagnetic particles and the physiological fluid are mixed in a vessel. Once the immunoreactive agents, i.e. antibody layer, has bonded with the target population cells or chemical species, the particles are separated from the fluid by activating a magnetic coil to capture and immobilize the particles and opening a valve to release the fluid from the vessel. The immunoreactive paramagnetic particles are then transferred to another vessel containing a cleaving agent. This cleaving agent promotes the release of the selected target population from the coated paramagnetic particles.
The above discussed procedures and apparatus allow for the separation of a target population, whether cells, proteins or antibodies, from a physiological fluid. The procedures for which these methods are used are typically those in which it is desired to remove this target population from the physiological fluid prior to returning the fluid to the patient. For example, the above procedure is useful for removing infected or tumor cells from a specific tissue, e.g. bone marrow. The major drawback to the described procedures and apparatus is the complexity of the procedure or apparatus used, or the inability of thoroughly separating the target population from the remainder of the physiological fluid. That is, the disclosed procedures and apparatus were not as concerned with the complete removal of most of the physiological fluid since the primary focus is the removal of the target population. Thus there remains the risk of some contamination of the target population with extraneous cells or compounds from the fluid.
Further, such procedures and apparatus have the drawback of requiring multiple containers for performing the isolation, as described in Giaever et al, or require specific knowledge of procedures to insure prevention of contamination of the final product. It thus remains desirable to provide for a method and apparatus which would simplify the handling and isolation of a target population, and specifically an antibody, for the purpose of easily, and safely preparing a pharmaceutical composition.